ArciTect™
ArciTect™ CRISPR-Cas9 Genome Editing System for Cell Biologists
The ArciTect™ product family is a CRISPR-Cas9 genome editing system that uses ribonucleoprotein (RNP) complexes composed of purified Cas9 protein and custom synthetic guide RNA. Compared to previous technologies that utilize plasmid or mRNA-based systems, an RNP-based system enables efficient delivery and expression of CRISPR machinery in difficult-to-manipulate cells, including stem and primary cells. Once inside the cell, the RNP complexes will not induce the cellular immune response and will degrade in a timely manner to reduce off-target effects (Table 1). With validated reagents and protocols, the ArciTect™ system enables you to perform high-efficiency genome editing and generate functional gene-edited cells in your own lab.
Overcome Hurdles in Efficiency
Overcome challenges in efficient delivery and expression of CRISPR machinery using an RNP-based CRISPR-Cas9 system. ArciTect™ enables researchers to perform high-efficiency genome editing in difficult-to-manipulate cell types, including stem and primary cells.
Why Use ArciTect™?
- Maximize delivery and expression in difficult-to-manipulate cell types by using RNP complexes.
- Simplify genome editing with an integrated guide RNA design tool and cell-type-specific protocols.
- Get to your results faster with ready-to-use purified Cas9 proteins and synthetic guide RNAs.
- Minimize potential off-target cutting with timely degradation of the RNP complex.
Figure 1. Genome Editing Workflow
Table 1. Comparison Between Different CRISPR Methods. 1
Genome Editing Protocols and Data
Explore step-by-step instructions for performing high-efficiency genome editing using CRISPR-Cas9 in a variety of cell lines, including stem and primary cell types. The protocols are optimized and validated with a case study and supporting data, and include important cell culture considerations and methods to evaluate editing efficiency.
Human Pluripotent Stem Cells (hPSCs)
Gene knockout and knock-in in hPSCs using electroporation and chemical transfection.
Case study: Knockout and Knock-in optimization through GFP to BFP conversion
Human Primary T Cells
Gene knockout in primary human T cells using electroporation.
Case study: Evaluation of optimal culture methods for high-efficiency TRAC knockout
CD34+ Human Hematopoietic Stem and Progenitor Cells (HSPCs)
Gene knockout in HSPCs using electroporation.
Case study: Evaluation of optimal culture methods for high-efficiency genome editing in CD34+ HSPCs
Human Intestinal Organoids
Gene knockout in human adult stem cell-derived intestinal organoids using electroporation.
Genome Editing Products
Guide RNA (gRNA)
Features:
- Single guide RNA (sgRNA) containing the crRNA and tracrRNA regions within a single molecule
- Two-part gRNA system composed of ArciTect™ crRNA and ArciTect™ tracrRNA
- Compatible with all Cas9 nucleases
Applications:
- Use ArciTect™ sgRNA or ArciTect™ crRNA and tracrRNA to guide the Cas9 nuclease to a specific target location
Cas9
Features:
- RNP complex is active immediately following transfection
- All versions of Cas9 contain nuclear localization signals for rapid translocation into the nucleus
Applications:
- Use ArciTect™ Cas9 Nuclease to generate double-strand breaks at specific locations in the genome
- Use ArciTect™ Cas9-eGFP Nuclease to optimize transfection conditions or sort cells following transfection
Human HPRT
Features:
- Validated for use with ArciTect™ family of products for genome editing
Applications:
- Use ArtiTect™ Human HPRT crRNA and Primer Mix as a positive control to optimize transfection protocols
- ArciTect™ Human HPRT Primer Mix can be used in a T7 endonuclease assay to assess cleavage efficiency
Scientific Resources
Wiley E-book: Genome Editing Applications
Learn about next-generation disease modeling using CRISPR, including comprehensive genome editing strategies for complex cell culture models, suggestions for optimizing experimental conditions, and troubleshooting tips.
References
- Liang X et al. (2015) Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection. J Biotechnol. 208: 44-53.